LOCUS       V027235                 8865 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027235
VERSION     V027235
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3545
                     /label="Adar(NM_001410659)"
                     /note="Adar(NM_001410659)"
                     /gene="Adar"
     CDS             3552..3617
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3641..4229
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4295..4343
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4545..4973
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4987..5316
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5383..6177
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6353..6474
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6523..6539)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6547..6563
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6571..6601)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6616..6637
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6925..7513)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7684..8544)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8545..8649)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc tgaaatcaag gagaagatct
      781 gtgactatct gttcaatgtg tcaaactcct ctgccctgaa cctggctaag aacattggcc
      841 tcaccaaggc ccgagatgtg acctcagtgc tgattgactt ggaaaggcaa ggcgatgtct
      901 acaggcaagg ggcaactcct cccatctggt acttgacgga caagaagcgt gagaggctgc
      961 agatgaagag aagtacacac agtgctcctg cccctacccc gacagctgtc ccagaggcca
     1021 ctagaagccc ctcattccct gcctgccacc cgcccccagc aggtgcctca agcagtgtgg
     1081 cagcctccaa gagagtggag aatgggcagg agcctgcgat aaagcatgaa agtaggcatg
     1141 aggccagacc aggaccaatg agactgcggc ctcacgctta tcacaatggc ccctctagag
     1201 cagggtatgt ggcctctgaa aatggccagt gggccacaga tgacatccca gataacttga
     1261 atagtatcca cacagcacca ggtgagtttc gagccatcat ggagatgccc tccttctaca
     1321 gccctacctt gccacggtgt tcaccctaca agaagctaac tgagtgccag ctgaagaacc
     1381 ctgtcagcgg gttgttagag tatgctcagt tcactagtca gacctgtgat ttcaacctga
     1441 tagagcagag tggaccgtcc catgaacctc gatttaaatt ccaggttgtc atcaatgggc
     1501 gggaatttcc cccagctgag gctggcagca agaaagtagc caagcaggac gcagcagtga
     1561 aagccatggc gattctgctt cgggaagcca aagccaaaga cagtggtcaa ccagaagact
     1621 tgtcccactg tcccatggaa gaagactcgg agaaaccagc agaggctcag gcccccagct
     1681 cctcagcaac atccttgttc tctgggaaga gcccagttac tacactgctt gagtgcatgc
     1741 acaaactagg gaactcctgt gaattccgtc tcctgtccaa agaaggccct gctcatgacc
     1801 ccaagttcca gtactgtgta gcagtaggag cccagacctt cccccctgtg agcgccccca
     1861 gcaagaaggt agcaaagcag atggccgcag aggaagccat gaaggcgctg caggaggagg
     1921 cagccagttc agcggatgac cagtctggag gtgcgaacac agactcactt gatgaatcta
     1981 tggctcccaa caagatcagg aggattggtg agctcgtcag gtacctgaac accaaccccg
     2041 taggcggctt gttggagtac gcccgatctc atggctttgc tgctgagttc aagctcattg
     2101 accagtctgg acctcctcac gaacccaagt ttgtttacca agcaaaagtt gggggccgct
     2161 ggtttccagc cgtgtgtgca cacagcaaga aacagggcaa gcaggatgca gcggatgcag
     2221 ccctccgggt cttgatcggg gagagcgaga aggcagagca gttgggtttc gcagaggtaa
     2281 ccccagtaac aggggccagt ctcagaagaa ctatgctcct cctttccagg tccccagatg
     2341 cacatccaaa gacacttcct ctctctggca gcaccttcca cgaccagata gctatgctga
     2401 gccacaggtg cttcaatgct ctgaccaaca gtttccagcc ctccctgctc ggccgcaaga
     2461 tcctggctgc cattattatg aaaagagatc cagaggacat gggtgttgtc gtgagtttgg
     2521 ggacaggcaa tcgctgtgtg aaaggggact ctctcagcct gaagggagag acggtcaatg
     2581 actgccatgc cgaaatcatc tcccggaggg gcttcatcag gtttctctac agtgaactga
     2641 tgaagtacaa ccaccacact gccaagaaca gcatatttga gcttgccagg ggaggagaga
     2701 agctgcagat aaagaagacg gtttcttttc atctctacat cagcacggcg ccatgtggag
     2761 atggggccct ctttgacaaa tcctgcagtg accgtgccgt ggaaagcaca gagtcccgcc
     2821 attaccctgt ctttgaaaat cccaagcaag gcaagcttcg caccaaggtg gagaatgggg
     2881 aaggcacaat tcctgtggag tccagtgata ttgtacccac gtgggatggc atccgccttg
     2941 gggaaagact ccgtaccatg tcctgtagtg acaaaatcct acgctggaat gtgctgggcc
     3001 tgcaaggggc gttgttgacg cacttcctac agcctgtgta cctgaaatct gtaaccttag
     3061 gttacctttt cagccaaggg catctgaccc gtgctatttg ctgccgcgtg accagagatg
     3121 ggaaagcatt tgaggatgga ctacgctatc cctttattgt caaccacccc aaggtcggcc
     3181 gagtcagtgt ttatgattcc aaaagacagt ccggaaagac caaggaaaca agcgtcaact
     3241 ggtgcatggc tgatggctat gacctagaga tcctggatgg caccagaggc actgtggatg
     3301 gaccagggaa agagttgtct cgggtgtcca agaagaatat tttccttcag tttaagaagc
     3361 tctgctcctt ccgagcccgc agagatttac tgcagctctc ttatggtgaa gccaagaaag
     3421 ctgcccgtga ctatgactta gccaagaact acttcaagaa aagcctgcga gacatgggct
     3481 atgggaattg gatcagcaaa ccccaggagg aaaagaactt ttacctctgt ccagtaccca
     3541 atgacggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     3601 aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa
     3661 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3721 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3781 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3841 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3901 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3961 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     4021 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     4081 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     4141 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     4201 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     4261 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4321 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4381 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4441 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4501 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4561 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4621 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4681 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     4741 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4801 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4861 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     4921 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4981 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5041 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5101 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5161 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5221 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     5281 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     5341 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5401 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5461 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5521 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5581 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5641 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5701 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5761 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5821 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5881 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5941 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     6001 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     6061 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6121 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6181 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     6241 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6301 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     6361 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6421 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6481 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6541 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6601 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6661 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6721 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6781 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6841 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6901 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6961 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7021 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7081 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     7141 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7201 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7261 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7321 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7381 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7441 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7501 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7561 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7621 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7681 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7741 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7801 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7861 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7921 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7981 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8041 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8101 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8161 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8221 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8281 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8341 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8401 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8461 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8521 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8581 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8641 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     8701 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     8761 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     8821 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
